Δ3-Δ2-Enoyl-CoA isomerase from the yeast Saccharomyces cerevisiae: Molecular and structural characterization
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Chapter 4. Materials and methods

Table of Contents
4.1. Strains and plasmids
4.2. Saccharomyces genome database
4.3. Gene disruption (I)
4.4. Analysis of transcriptional regulation (I)
4.5. Subcellular localization (I)
4.6. Construction of the ECI1 expression vector (I)
4.7. Expression of the recombinant protein (I)
4.8. Protein purification (I, II)
4.9. Enzyme assays (I)
4.10. Site-directed mutagenesis (III)
4.11. Determination of native molecular mass (I, III, IV)
4.12. Crystallization (II, III, IV)
4.13. X-ray diffraction data collection and processing (II-IV)
4.14. Structure determination by the MAD method and refinement of the structure (III)
4.15. Structure determination by the molecular replacement method and refinement of the structure (IV)
4.16. Structure analysis and validation (III-IV)
4.17. Comparison of the structures belonging to the hydratase/isomerase superfamily (IV)

Most of the methods are described in more detail in the original articles referred to by their Roman numerals.

4.1. Strains and plasmids

All the bacterial and yeast strains as well as the construction of the plasmids used in the experiments are described in the original articles.

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Aims of the present study Saccharomyces genome database

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